Abscission is the ability of plants to slough off organs, such as fruit, by an active separation of cells. Abscission occurs in specific areas of organs to be shed called xe2x80x9cabscission zones.xe2x80x9d Abscission zones contain several layers of starch-filled cells that eventually undergo striking changes such as cell wall digestion that ultimately lead to dropping of the fruit from the stem.
Commercial harvesting of fruit can require deviation from the natural abscission cycle. When harvesting takes place where the abscission layer has not begun to separate, a great deal of force can be required to remove the fruit. The force can damage the fruit or the plant. Presently available abscission agents, such as Ethrel(copyright), can cause unwanted defoliation of a plant or tree. Compositions and methods are needed in the art to reduce the unwanted defoliation of plants and trees, particularly where defoliation is caused by an abscission agent or by an herbicide.
It is an object of the invention to provide methods and compositions for reducing the defoliation of plants and trees. This and other objects of the invention are provided by one or more of the embodiments described below.
One embodiment of the invention provides a defoliation safener composition. The composition comprises an abscission agent, a thinning agent, or a combination thereof, and an xcex12-adrenergic receptor agonist, an adrenergic inhibitor, or a combination thereof.
Another embodiment of the invention provides a defoliation safener composition. The composition comprises a plant-compatible surfactant and an xcex12-adrenergic receptor agonist, an adrenergic inhibitor, or a combination thereof.
Yet another embodiment of the invention provides a defoliation safener composition. The composition comprises a chemical selected from the group consisting of an herbicide, a pesticide, an antimicrobial agent, a fungicide, a fertilizer, and a plant hormone; and an xcex12-adrenergic receptor agonist, an adrenergic inhibitor, or a combination thereof.
Still another embodiment of the invention provides a method of reducing defoliation of a plant or tree comprising administering an effective amount of an xcex12-adrenergic receptor agonist, an adrenergic inhibitor, or a combination thereof, to a plant or tree, whereby defoliation is reduced.
Therefore, the invention provides methods and compositions useful in the reduction of defoliation of plants and trees.
Adrenergic receptors (or adrenoceptors) mediate the central and peripheral actions of the primary sympathetic neurotransmitter, norepinephrine, and the primary adrenal medullary hormone, epinephrine. See, Ruffolo et al., Med. Res. Rev. 14:229 (1994). Adrenergic receptors mediate a variety of functions and have affinities for many synthetic drugs.
Adrenergic receptors are divided into two subtypes: xcex11-adrenergic receptors and xcex12-adrenergic receptors. xcex12-adrenergic receptors are located on presynaptic nerve terminals, postsynaptically, and in non-synaptic settings, such as human platelets, adipocytes, and vascular smooth muscle. Agonists of xcex12-adrenergic receptors are useful in treating a variety of diseases including, for example, hypertension, spasticity (resulting from stroke, cerebral trauma, or multiple sclerosis), short stature (in children with constitutional growth delay), glaucoma, gastric ulcers, pain relief, behavior disorders (e.g., attention-deficit hyperactivity disorder, conduct disorder, oppositional defiant disorder, Tourette""s syndrome, Lesch-Nyhan syndrome, and the disinhibitory symptoms accompanying post-traumatic stress disorder and dementia), and diarrhea associated with diabetes. See, e.g., U.S. Pat. No. 6,147,102; U.S. Pat. No. 5,854,290; U.S. Pat. No. 4,312,886.
Adrenergic inhibitors are also useful in the invention and include, for example, guanethidine, bethanidine, guanadrel, and debrisoquine.
It has now been discovered for the first time that agonists of xcex12-adrenergic receptors and adrenergic inhibitors and combinations thereof are useful as safeners, which reduce defoliation of plants and trees. In particular, agonists of xcex12-adrenergic receptors and adrenergic inhibitors are useful in reducing defoliation of plants and trees resulting from the administration of an abscission agent or herbicide to a plant or tree.
Any xcex12-adrenergic receptor agonist can be used in the invention. Preferably, the xcex12-adrenergic receptor agonist is clonidine, UK14,304 (Brimonidine), apraclonidine, guanfacine, guanabenz, phenylephrine, methoxamine, metaraminol, ephedrine, oxymetazoline, naphazoline, tetrahydrozoline, or analogues thereof. An analogue is structurally similar to a particular xcex12-adrenergic receptor agonist and has similar biological activity to the xcex12-adrenergic receptor agonist. Similar biological activity means having 5%, 10%, or 15% greater or lesser biological activity than the particular xcex12-adrenergic receptor agonist.
Furthermore, adrenergic inhibitors can be used in the invention. Preferably, the adrenergic inhibitor is guanethidine (Ismelin), bethanidine, guanadrel (Hylorel), debrisoquine, or analogues thereof.
An analogue is structurally similar to a particular xcex12-adrenergic receptor agonist or adrenergic inhibitor and has similar biological activity to the xcex12-adrenergic receptor agonist or adrenergic inhibitor. Similar biological activity means having 5%, 10%, or 15% greater or lesser biological activity than the particular xcex12-adrenergic receptor agonist or adrenergic inhibitor.
xcex12-adrenergic receptor agonists and adrenergic inhibitors are known in the art and are commercially available from suppliers such as Sigma/Aldrich (St. Louis, Mo.) and Tocris (Ballwin, Mo.).
Compositions of the invention can be applied to a plant or tree in a mixture with a carrier, or optionally, other auxiliary agents from any one of the standard types of preparations commonly used in agriculture, for example, a dry blend, granules, a wettable powder, an emulsion, and an aqueous solution. Suitable carriers for solid formulations include clay, talc, kaolin, bentonite, terra abla, calcium carbonate, diatomaceous earth, silica, synthetic calcium silicate, kieselguhr, dolomite, powdered magnesia, Fuller""s earth, gypsum and the like. Solid compositions can be in the form of dispersible powders or grains, comprising, in addition to an xcex12-adrenergic receptor agonist or adrenergic inhibitor, a surfactant to facilitate the dispersion of the powder or grains in liquid. Granular compositions can be prepared by, for example, impregnating an xcex12-adrenergic receptor agonist or adrenergic inhibitor onto or into granulated carriers such as attapulgites or vermiculites, or granulated solid fertilizers.
Liquid forms of the compositions of the invention, which are used for example, for spraying on plants or trees as aerosols or mists, are prepared for application by mixing an xcex12-adrenergic receptor agonist or adrenergic inhibitor, surfactant, if used, and a liquid carrier, such as water or other liquid to form a stable emulsion or suspension. Liquid compositions of the invention include solutions, dispersions, or emulsions containing an xcex12-adrenergic receptor agonist or adrenergic inhibitor and optionally, one or more surface-active agents such as wetting agents, dispersing agents, emulsifying agents, or suspending agents.
xcex12-adrenergic receptor agonist and adrenergic inhibitor compositions of the invention are preferably formulated for use in a mixture with a plant-compatible surfactant, to provide solid or liquid (including a suspension of a solid in a liquid phase) formulations. A plant-compatible surfactant causes little or no damage to plants or trees. Surfactants of the invention can be nonionic, anionic, cationic, or zwitterionic surfactants. A surfactant can be present in a composition of the invention as formulated or, alternatively, a surfactant can be introduced to the compositions of the invention during application to the plant or tree. Preferably, a surfactant is Kinetic(copyright) (proprietary blend of poloyalkylene oxide modified polydimethyl siloxane and non-ionic surfactant) (Setre Chemical Co., Memphis, Tenn.), Silwet L-77(copyright) (polyalkylene oxide modified heptamethyl trisiloxane) (OSi Specialties, Danbury Conn.), or Tween(copyright) 20 (sorbitan mono-9 octadecenoate poly(oxy-1,1-ethanedlyl)) (Sigma-Aldrich, St. Louis Mo.). Preferably, Kinetic(copyright) and Silwet L-77(copyright) are used at a final concentration of about 1.0 to 0.01%. More preferably, Kinetic(copyright) and Silwet L-77(copyright) are used at a final concentration of about 0.5 to 0.05 %. Even more preferably, Kinetic(copyright) and Silwet L-77(copyright) are used at a final concentration of about 0.15%. Preferably, Tween(copyright) 20 is used at a final concentration of about 1.0 to 0.01%. More preferably, Tween(copyright) 20 is used at a final concentration of about 0.5 to 0.05%. Even more preferably, Tween(copyright) 20 is used at a final concentration of about 0.1%.
xcex12-adrenergic receptor agonist and adrenergic inhibitor compositions of the invention can be used alone or in a mixture or combination with abscission agents, thinning agents, plant hormones, fertilizers, pesticides, herbicides, antimicrobial agents, or fungicides. Preferably, the xcex12-adrenergic receptor agonist and adrenergic inhibitor compositions of the invention are used in combination or mixture with sulfonylureas and imidazolinones. Even more preferably, the xcex12-adrenergic receptor agonist and adrenergic inhibitor compositions of the invention are used in combination or mixture with metsulfuron-methyl, an herbicide commonly used to control broadleaf weeds in agronomic crops. Additionally, the xcex12-adrenergic receptor agonist and adrenergic inhibitor compositions of the invention are especially useful in combination or mixture with an abscission agent such as Ethephon(copyright) or Ethrel(copyright).
Compositions of the invention can also be used in plant substantive formulations. Plant substantive formulations mechanically or chemically adhere to a plant and resist removal. Preferred plant substantive agents include various waxes and paraffins, polymers, and sulfur. Wax coated compositions can be prepared by dispersing an xcex12-adrenergic receptor agonist or adrenergic inhibitor or combinations thereof in molten wax, forming the dispersion into small particles, and cooling the composition below the melting point of the wax. The water resistance of the particles can be controlled by increasing or decreasing the amount of wax employed so as to provide proper release for climatic conditions, i.e., wet areas or dry areas. Additionally, various additives can be dissolved in the wax phase in order to improve the water resistance of the composition or effect other benefits such as slow-release additives or anticaking agents. Other plant substantive agents include casein, salts of alginic acids, cellulose gums and their derivatives, polyvinyl pyrrolidone, vegetable gums, propylene glycol, invert syrup, corn syrup, and the like.
xcex12-adrenergic receptor agonists, adrenergic inhibitors, or combinations thereof are administered to any type of monocotyledon or dicotyledon plant or tree to reduce unwanted defoliation, gummosis, or twig-dieback. A defoliation safener composition of the invention reduces unwanted defoliation, gummosis, or twig-dieback, or a combination thereof. Unwanted defoliation, gummosis, or twig-dieback can be caused by, for example, administration of an abscission agent, a thinning agent, a herbicide, a plant hormone, a fertilizer, a pesticide, an antimicrobial agent, or a fungicide to a plant or tree. Unwanted defoliation, gummosis, or twig-dieback can also occur due to natural causes, including, for example, adverse climatic conditions. The compositions of the invention can be administered to a whole plant or tree or to a portion of a plant or tree. Administration of the compositions of the invention can occur once or can occur two or more times, where each administration is separated by a time interval, such as about 1, 3, 5, or 10 days.
A composition of the invention can be used in combination with an abscission agent or a thinning agent to reduce unwanted defoliation, gummosis, or twig-dieback associated with the agent. Administration of the compositions can occur before, during, or after the application of an abscission or thinning agent. For example, the compositions of the invention can be administered about 1, 3, 5, or 10 days before or after the administration of an abscission or thinning agent. Abscission agents include, for example, Ethrel(copyright) (2-chloroethyl phosphonic acid) (Aventis), Ethephon6(copyright) (Micro Flo Co., Memphis, Tenn.), Harvade(copyright) (Uniroyal Chemical Co., Middlebury, Conn.), FreeFall(copyright) (Griffin LLC, Valdosta, Ga.), and Folex(copyright) (Aventis). Thinning agents include, for example, naphthanlene acetic acid and Accel(copyright) (6-benzyladenine).
In another embodiment of the invention, an xcex12-adrenergic receptor agonist, adrenergic inhibitor, or combinations thereof are combined with an abscission agent comprising corotanine. Corotanine (COR) is a polyketide that is produced by several pathovars of Pseudomonas syringae. See Bender et al., Microbiol. Mol. Biol. Rev. 63:266-292 (1999). COR is a phytotoxin comprised of coronafacic acid (CFA), a polyketide, and coronamic acid (CMA), an ethylcyclopropyl amino acid. COR does not act as a phytotoxin on trees and plants of the invention, and in particular does not act as a phytotoxin to citrus.
COR is produced by chemical synthesis or by fermentation of cultures of microorganisms that produce COR, either naturally or recombinantly. For example, COR can be produced in vitro by P. syringae strain PG4180.N9 or other high coronatine-producing strain. A starter culture is prepared by making 10 mL of broth containing 10 g/L mannitol, 2 g/L L-glutamic acid, 0.5 g/L KH2PO4, 0.2 g/L NaCl, 0.2 g/L MgSO4, 0.35 g/L yeast extract, and 10 xcexcg/mL kanamycin, pH 7.0. A high coronatine-producing P. syringae strain is inoculated into the broth and the starting culture is incubated for 48 hours in an incubator-shaker operating at 200 rpm and 28xc2x0 C.
A fermentation culture is prepared by making a 1 L broth solution containing 450 mL HSC medium A (1 g/L NH4Cl, 0.2 g/l MgSO4, 4.1 g/L KH2PO4, 0.3 g/L KNO3, and 10 mL of a solution of 2 mM FeCl3, pH 6.8), 50 mL HSC medium B (20% glucose), and 10 xcexcg/mL kanamycin. 25 mL of a starting culture is added to the fermentation culture and it is incubated for 6 days at 18xc2x0 C. in an incubator-shaker operating at 200 rpm.
The fermentation culture is centrifuged for 20 minutes at 5,000xc3x97 g at 4xc2x0 C. and the pellet is discarded. The pH of the supernatant is adjusted to 2.5 with 1 N HCl. The supernatant containing coronatine is as partitioned three times against ethyl acetate (120 mL ethyl acetate: 100 mL supernatant) and the aqueous phases are discarded. The organic phases are combined and concentrated to 15 mL under vacuum at 35xc2x0 C. The concentrate is partitioned three times against a solution of 50 mM sodium carbonate, pH 10.5 (15 mL sodium carbonate solution:15 mL concentrate) and the organic phases are discarded. The aqueous phases are combined and the pH is adjusted to 2.5 with 1 N HCl. The aqueous phase is partitioned three times against ethyl acetate (15 mL aqueous:15 mL ethyl acetate). The aqueous phases are discarded and the organic phases are combined. Excess water from the organic phase is removed by adding 5 g anhydrous sodium sulfate and gently agitating. The organic phase is decanted and evaporated to dryness under vacuum at 35xc2x0 C. The coronatine is resuspended in 20 mL methanol. 0.5 mL of the suspension can be removed for quantitation using HPLC as described in Palmer and Bender (Appl. Environ. Microbiol. 59:1619-1626 (1993)). The remainder of the suspended coronatine can be evaporated to dryness and stored at xe2x88x9280xc2x0 C., if necessary. Typically, 100 mL of starter culture yields 30 mg of coronatine.
Any fermentation technique, can be used to produce COR, including, for example, batch, fed-batch, semi-batch, or continuous fermentation.
A composition of the invention can also be used in combination with chemicals such as herbicides, plant hormones, fertilizers, pesticides, antimicrobial agents, and fungicides to reduce unwanted defoliation, gummosis, and twig-dieback associated with the administration of the chemicals. Administration of the compositions can occur before, during, or after the application of a chemical.
In one embodiment of the invention, the compositions of the invention along with an abscission agent can be administered to a fruit-bearing plant or tree. The compositions of the invention can be administered in a mixture or combination with an abscission agent or can be administered before or after the administration of an abscission agent. When fruit has reached substantial maturity, a composition of the invention is applied along with an abscission agent, generally about 3 to 20 days prior to harvest, and preferably about 3 to 10 days prior to the harvesting date by administration to the fruit locus or to the whole plant or tree. Optionally, two or more applications of compositions of the invention can be done at different times prior to harvest. Preferably, application is by spray, to run-off.
Preferably, the concentration of an xcex12-adrenergic receptor agonist, adrenergic inhibitor, or combination thereof is about 25 to 1,000 mg/L (on a weight/volume basis). More preferably, the concentration is about 250 xcexcM (62 mg/L) to 2.0 mM (500 mg/L). Even more preferably, the concentration is about 50 mg/L to 500 mg/L. For citrus trees, approximately 2.5 liters of 250 xcexcM to 2.0 mM is used per fifteen foot tree.
Fruit bearing plants or trees of the invention include the major types of fruit for example, berries such as grapes, blueberries, and oranges; drupes such as peaches, cherries, olives, plums and walnuts; aggregate fruit such as blackberries and raspberries; multiple fruit such as pineapples, figs and mulberries; and accessory fruit such as apples, pears and strawberries. Also included are cotton plants. Preferred fruit of the invention are citrus fruit such as oranges, grapefruit, kumquats, lemons, limes, tangerines, temples, citrange, tangelo, pomelo, and citron.
The following are provided for exemplification purposes only and are not intended to limit the scope of the invention described in broad terms above. All references cited in this disclosure are incorporated herein by reference.